PROJECT SUMMARY Ethanol consumption contributes to approximately 6% of global deaths and is a major cause of morbidity and mortality in the United States. According to NIAAA statistics, about 57% of adults report consuming alcohol recently and 25% of adults engage in binge drinking. These statistics support a pressing need for understanding the biochemical mechanisms underlying chronic and binge drinking toward the pathogenesis of alcoholic liver disease. Ethanol metabolism is known to alter numerous biochemical pathways, including glycolysis, lipid metabolism, and the TCA cycle, as well as the induction of oxidative stress. The overall concept of oxidative damage in ethanol toxicity is well known, but altered thiol redox signaling and control is an area of investigation largely underexplored in ethanol toxicity and has major implications for multiple pathologic processes, including inflammation, cell growth, and metabolism. Therefore, we present an innovative approach for investigating how alcohol-mediated induction of hepatic mitochondrial protein acetylation impacts thiol redox signaling and control. We will investigate the proposed specific aims: Specific Aim 1: Investigate the metabolic impact of acute ethanol toxicity on hepatic mitochondrial thiol redox signaling and control. Specific Aim 2: Understand the impact of chronic ethanol metabolism on hepatic mitochondrial thiol redox signaling and control. Specific Aim 3: Characterize the regulatory implications of ethanol metabolism on protein thiol switches. These aims will be interrogated utilizing a multidisciplinary approach and cutting-edge targeted and untargeted ?OMICS techniques, as well as innovative data analyses and bioinformatics. Additionally, computational integration of multiple different ?OMICS datasets with other biomarkers of oxidative stress and liver damage represent a substantial intellectual innovation that will provide previously undiscovered mechanistic leads that will fuel future research aims. Lastly, understanding the mechanisms by which ethanol metabolism impacts hepatic thiol redox systems will support the development of novel therapeutic strategies to ameliorate alcoholic liver disease in millions of Americans.